1. Field of the Invention
The invention relates to methods, kits, and systems for detection of a plurality biological agents in a sample utilizing a minimal number of containers, useful for diagnostic assays and screening for disease agents.
2. Description of the Related Art
Diagnostic assays that sensitively, specifically, and quickly detect biological agents, e.g., pathogens, in samples are becoming increasingly important for both disease and diagnostic bioagent monitoring. Few assays are able to accurately detect physiologically or clinically relevant organisms on an appropriate time-scale for the early detection of the presence of an infective or otherwise harmful agent. To date, the most sensitive detection methods involve PCR. Determining the presence or absence of a plurality of biological agents in a single sample can be performed using multiplexed detection methods. Multiplexed real time PCR is one method that can be used for this type of diagnostic assay.
Assays based on PCR can be limited by the complexity of optimizing the PCR reactions to test for multiple agents in a cost-effective number of reaction tubes. As a general rule, the number of probes needed to support a highly specific confirmation result range from two to as many as six sequences. As one of skill in the art will be aware, optimizing a PCR reaction with many different primer pairs and probes can be a formidable task that becomes increasingly unmanageable as the number of agents to be detected increases.
Assays based on PCR can also be limited by the number of unique chemical labels available for analysis of results. For example, real time PCR assays generally employ fluorescent labels. When performing multiplexed real time PCR, the number of labels that can be used in a single reaction is limited by the number of fluorescent color channels available in the optical detection system used.
An attractive approach to overcome these and other limitations of current multiplex assays is provided by the present invention.